Overview

  • Product name
  • Description
    Rabbit polyclonal to SM22 alpha
  • Host species
    Rabbit
  • Specificity
    Recognises a band at 23kD that is specifically blocked by the immunising peptide.
  • Tested applications
    Suitable for: ICC/IF, IHC-P, IHC-Fr, WB, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human, Pig
  • Immunogen

    Synthetic peptide within Mouse SM22 alpha aa 150 to the C-terminus conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab16067)

  • Positive control
    • WB: Human smooth muscle cell lysate. HeLa whole cell and nuclear extract. Human skeletal muscle tissue lysate. Human, mouse and rat colon tissue lysate. ICC/IF: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab14106 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 - 5 µg/ml.
IHC-P 1/200.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ICC Use at an assay dependent concentration.

Target

Images

  • All lanes : Anti-SM22 alpha antibody (ab14106) at 1/1000 dilution

    All lanes : Lysates prepared from human primary smooth muscle cells

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated bovine polyclonal to rabbit IgG at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 23 kDa


    Exposure time: 2 minutes

    See Abreview

  • ab14106, at 1/100, staining SM22 alpha in chicken smooth muscle tissue sections by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections).

    Sections were fixed in methacarn (60% methanol, 30% chloroform, 10% acetic acid), prior to blocking in 2% serum for 5 minutes at 25°C and then incubated with ab14106, for 1 hour at 25°C. An Alexa-Fluor® 488 goat polyclonal to rabbit Ig, diluted 1/100, was used as the secondary antibody.

    See Abreview

  • ab14106 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed in 100% methanol for 5 minutes, permeabilized in 0.1% PBS-Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14106 at 1 µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed used at a 1/1000 dilution for 1 hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature.

    DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1 hour at room temperature.

  • ab14106 staining SM22 alpha in mouse muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% serum for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/1000.

    See Abreview

  • All lanes : Anti-SM22 alpha antibody (ab14106) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa nuclear extract
    Lane 3 : Human skeletal muscle tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

    Predicted band size: 23 kDa
    Observed band size: 23 kDa



    The band was completed abolished by peptide blocking with ab16067 (immunizing peptide) - not shown.

     


     

     

     

  • All lanes : Anti-SM22 alpha antibody (ab14106) at 1 µg/ml

    Lane 1 : Human colon tissue lysate - total protein (ab30051)
    Lane 2 : Mouse colon tissue lysate
    Lane 3 : Rat colon tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 23 kDa
    Additional bands at: 115 kDa, 20 kDa, 36 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 5 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab14106 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Anti-SM22 alpha antibody (ab14106) at 1 µg/ml + Recombinant Human SM22 alpha protein (ab101469) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa


    Exposure time: 30 seconds

References

This product has been referenced in:
See all 166 Publications for this product

Customer reviews and Q&As

1-10 of 36 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse aortic smooth muscle cells)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
25 µg
Treatment
Cre or GFP adenovirus 6 hrs
Specification
Mouse aortic smooth muscle cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Mar 15 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (vascular smooth muscle cells)
Permeabilization
No
Specification
vascular smooth muscle cells
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 15 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Carotid artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: sodium citrate solution
Permeabilization
Yes - 0.3% Triton X-100/PBS
Specification
Carotid artery
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 15 2018

Application
IHC - Wholemount
Sample
Mouse Tissue (artery)
Specification
artery

Abcam user community

Verified customer

Submitted Mar 01 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Adult Heart)
Permeabilization
Yes - 0.5% Tween-20 in block
Specification
Adult Heart
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 03 2017

Application
Western blot
Sample
Cow Tissue lysate - whole (Kidney)
Gel Running Conditions
Reduced Denaturing (14)
Loading amount
20 µg
Specification
Kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Sep 28 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (vascular smooth muscle cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
vascular smooth muscle cell
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

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Verified customer

Submitted May 05 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human arterial myocytes)
Specification
Human arterial myocytes
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 08 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Cell (muscle cell)
Specification
muscle cell
Permeabilization
Yes - 0.1%Triton X-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 25 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
PBS, 1% BSA, 10% goat serum, 0.1% TX-100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample
Mouse Cell (primary embryonic epicardial)
Specification
primary embryonic epicardial
Permeabilization
Yes - 0.5% Triton X-100
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 30 2014

1-10 of 36 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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